African Journal of Biotechnology Vol. 12(7), pp.

735-743, 13 February, 2013

Available online at http://www.academicjournals.org/AJB
DOI: 10.5897/AJB10.1728
ISSN 1684–5315 ©2013 Academic Journals

Full Length Research Paper

Chemical evaluation of protein quality and phenolic

compound levels of some Cucurbitaceae oilseeds from
Cameroon
Mercy Bih Achu1*, Elie Fokou1, Germain Kansci1 and Martin Fotso2
1
Department of Biochemistry, Faculty of Science, University of Yaoundé I, Cameroon.
2
Centre for Research in Food and Nutrition, IMPM, Yaoundé, Cameroon.
Accepted 7 July, 2011

This work studies the chemical evaluation of protein quality and phenolic compound contents of some
Cucurbitaceae (egusi) oilseeds from different areas in Cameroon. These seeds are Cucumeropsis
mannii, Cucurbita maxima, Cucurbita moschata, Lagenaria siceraria and Cucumis sativus. The seeds
were cleaned, dried, ground and part of the powder was defatted. The defatted cakes were analysed for
total and soluble nitrogen, true proteins and amino acids, while the undefatted seeds were analysed for
phenolic compound contents. The defatted cakes had high total protein contents. The trichloroacetic
acid soluble fraction of these proteins ranged from 25% (C. maxima from North West) to 94% of total
proteins (C. sativus from Adamawa and South West), due to the post harvest treatment of the seeds.
They were rich in most essential amino acids, giving protein digestibility, corrected amino acid scores
of 0.67 for C. sativus and 0.48 for C. mannii which was for lysine, indicating that in the absence of
tryptophan and methionine, lysine was the limiting amino acid in these seeds. These seeds had low
levels of phenolic compounds (0.34 to 0.43%). Defatted C. mannii could be good for preparing infant
formula, especially when mixed with soybean, in order to increase its lysine content.

Key words: Cucurbitaceae (egusi) seeds, proteins, amino acids, phenolic compounds.

INTRODUCTION

There is an increasing prevalence of nutrition related free kernel meals of different Cucurbit species had 60 to
illnesses especially in Africa due to poverty and 70% of proteins. Jacks (1986) reviewed the usefulness of
insufficient knowledge on the nutritional and economic Cucurbit seeds and showed that globulins account for 70
importance of locally available and easily accessible to 90% of the protein, they are rich in arginine, aspartic
foods and foodstuffs. Studies on Cucurbitaceae seeds and glutamic acids, but deficient in lysine and sulfur-
have shown that they contain high protein levels with high containing amino acids, and that supplementation with
levels of most essential amino acids except lysine the limiting amino acids increases the values. Previous
(Sharma et al., 1986; Nwokolo and Sim, 1987; Martin, studies on the nutritive value of these Cucurbitaceae
1998; El-Adawy and Taha, 2001). High protein levels in oilseeds from different regions in Cameroon by Achu et
Cucurbit seeds have also been shown by other al. (2005) showed that these seeds and their defatted
researchers from various countries, such as Lal et al. cakes are rich in proteins (28 to 40.49% and 61 to
(1983) who studied the kernel oils of 15 species of 73.59% respectively). Kanar et al. (2006) investigated
Cucurbitaceae seeds from India and showed that the oil- some nutritional and antinutritional characteristics of
Cucumis sativus and Lagenaria vulgaris seeds and
showed that they contained 31.2 to 31.8% crude proteins
and that heat treatment reduced the trypsin inhibitor and
*Corresponding author. E-mail: [email protected]. Tel: +237 lectin activities in both samples to negligible levels. The
77 98 98 51. essential amino acid profile compared well with the
736 Afr. J. Biotechnol.

FAO/WHO scoring pattern except for a deficiency of phenolic compounds and some quality parameters of
lysine and isoleucine, with lysine being the first limiting pumpkin seed oil and showed that the total phenolics
amino acid in both seeds. Yanty et al. (2007) showed that content ranged from 24 to 50.93 mg GAE/kg of oil. The
the crude protein content of Cucumis melo seeds from individual phenolics were tyrosol, vanillic acid, vanillin,
Malysia was 25%. Loukou et al. (2007) also found similar luteolin and sinapic acid. The maximum antioxidant
results with Cucurbit seeds from Côte d’Ivoire where they capacity measured by the reduction of 2,2-diphenyl-1-
found that these seeds have protein levels ranging from picrylhydrazyl (DPPH) radical was 62%, which was
29 to 36%. Ojieh et al. (2008) continued to show that comparable to 0.16 mM Trolox equivalent (DPPH is a
Citrullus lanatus has a crude protein content of 23.4% stable radical that is used to screen free-radical-
with good quantities (g/100 g protein) of arginine (9.0), scavenging ability of compounds). Parry et al. (2006)
isoleucine (4.8), leucine (4.2), and phenylalanine (3.2) characterized cold- pressed onion, parsley, cardamom,
which are essential amino acids as well as glutamic acid mullein, roasted pumpkin, and milk thistle seed oils and
(16.9) and aspartic acid (16.3) and that “egusi” melon found that the seed oils may serve as dietary sources of
compares favourably with the known protein rich foods special fatty acids, tocopherols, carotenoids, phenolic
such as soybean, cowpeas, pigeon peas and pumpkin. compounds, and natural antioxidants.
Mariod et al. (2009) found that the protein content from Phenolic compounds and phytochemicals in food have
six Sudanese Cucurbit seeds ranged from 14 to 17.5%. been shown to have both adverse (antinutrients) and
These results show a great variability on the protein beneficial health effects (antioxidants) in humans. For
contents which depend on the specie and which also example, tannins exhibit their toxicity effects, by forming
seem to depend on the regions, as seen from the low protein-tannin complexes through multiple hydrogen
values obtained for Sudanese seeds. binding between their hydroxyl groups and the carboxyl
Phenolic compounds have been shown to have a lot of groups of protein peptide bonds of proteolytic enzymes in
beneficial effects as antioxidants, antithrombotic and anti- the gastrointestinal tract (Bressani et al., 1983). Although
inflammatory effects, and inhibit carcinogenesis (Kris- some saponins have been shown to be highly toxic under
Ertherton et al., 2002). Siger et al. (2008) investigated the experimental conditions, acute poisoning is relatively rare
content and antioxidant activity of phenolic compounds in both in animals and man (Osagie and Eka, 1988). Phytic
cold-pressed plant oils in Poland and showed that acid, lectins, tannins, saponins, enzyme inhibitors,
pumpkin (Cucurbita pepo L.) oils had the highest total cyanogenic glycosides and glucosinolates reduce the
phenolic content (2.5 mg/100 g) and phenolic acids availability of certain nutrients and impair growth. When
(vanillic acid; 11.4 and protocatechuic acid; 3.1 mg/100 g) used at low levels, phytic acid, lectins, enzyme inhibitors
and also displayed high antioxidant activity (70%). and saponins, reduce blood glucose and/or plasma
Xanthopoulou et al. (2009) studied the antioxidant and cholesterol and triacylglycerols levels. Phytic acid,
lipoxygenase inhibitory activities of pumpkin seed protease inhibitors, saponins, lignans and phytoestrogens
extracts and showed that most extracts tested have been shown to reduce cancer risks. Phenolic
demonstrated radical scavenging activity, with fractions compounds found in foods also contribute to their
rich in phenolics showing the highest activity and that the astringency, and may also reduce the availability of
presence of molecules being able to scavenge radicals certain minerals such as zinc. During thermal processing,
and inhibit lipoxygenase in pumpkin seeds may in part phenolic compounds may undergo oxidation and the
explain the health benefits attributed to them. Peričin et oxidized phenolics so formed, such as quinones, may
al. (2009) showed that phenolic acids in pumpkins (C. combine with amino acids, thus making them nutritionally
pepo) were caffeic acid (in the skin, oil cake meal and unavailable (Shahidi, 1997). The levels of some of these
dehulled kernel) and syringic acid (whole hull-less seed antinutrients are usually reduced during culinary
and dehulled kernel). The dominant phenolic compound transformations. A decline in phytic acid content was
was p-hydroxybenzoic acid amounting to 34.7% (hull-less achieved by dehydration of lentil (44%), white beans and
seed), 52.0% (oil cake meal), 51.4% (skin), 67.4% pink-mottled cream beans. Cooking and dehydration
(dehulled kernels) and 51.8% (hulls) of the total phenolic significantly reduced the levels of enzyme inhibitors and
acid content. Most phenolic acids were present in bound lectins in beans to negligible concentrations. Increased in
(esterified and insoluble) form, from 50.6% (skin) to vitro protein digestibility was produced by dehydration in
84.1% (hull-less seed). Edeoga et al. (2010) investigated these legumes from 12% to 15%. (Martín-Cabrejas et al.,
the pharmaceutical and therapeutic potential of some wild 2009).
Cucurbitaceae species (Lagenaria vulgaris, In view of the beneficial effects of proteins and phenolic
Trichosanthes cucumerina, Momordica charantia and compounds to health, it is important to discover new
Luffa cylindrical) from South - East Nigeria and showed sources of these compounds. There is still shortage of
that the seeds generally contained more alkaloids than data on the protein quality and the content in phenolic
the other parts (0.03 to 0.07 mg/ml). They contained compounds of these seeds grown in Cameroon, which is
more tannins (0.484 to 0.9 mg/ml) than the other essential in order to exploit these seeds for better health,
phytochemicals. Andjelkovic et al. (2010) studied the well-being and development. This work is therefore
 Achu et al. 737

aimed at evaluating the protein quality (nature of proteins The amino acid composition: This was analysed in C. mannii and
and amino acid composition) and total phenolic C. sativus seeds, for they are the most widely distributed (found in
most markets) and widely consumed seeds. 20 mg of defatted
compounds of five Cucurbitaceae seeds from Cameroon. sample were weighed into a dry tube and 5 ml of HCl (6n) added.
Norleucine was added to the sample at a final concentration of 0.25
mM as internal standard. The tubes were tightly corked (Teflon
MATERIALS AND METHODS cork) and put in an oven at 110°C for 24 h. This hydrolysed the
peptide bonds, liberating amino acids, but oxidized sulphur-
Collection and treatment of samples containing amino acids, cysteine and methionine. Tryptophan was
also destroyed by acid hydrolysis and so was not detected by this
The “egusi” samples were collected from different bio-climatic areas method. Asparagine and glutamine were also transformed into
in Cameroon. These regions are Sahel, High Savanna, Rain forest aspartic and glutamic acids respectively. After 24 h of hydrolysis, 50
and Swamp forest regions. The seeds; Cucumeropsis mannii, µl of the hydrolysate was collected (in duplicate) and dried under
(egusi melon), C. maxima, (pumpkin or squash gourd), Cucurbita vacuum. The samples were derived by phenylisothiocyanate (PITC
moschata, (musk melon), Lagenaria siceraria (bottle gourd or or Edman’s reagent) in order for the amino acids to be detected
calabash) and C. sativus, (“Ibo” egusi); were bought already dried spectrophotometrically at 254 nm. Since the reaction with PITC
under local conditions by the farmers. Where the seeds could not occurred at basic pH, the samples were first neutralized in a mixture
be found, the fruits were bought from the farmers during periods of of ethanol/water/triethylamine (TEA) (2/2/1 v/v/v). 50 µl of this
harvest and the seeds extracted, washed, sun-dried and mixture was added in the tube and dried under vacuum. For
decorticated. derivation, 40 µl of a mixture of ethanol/water/TEA/PITC (7/1/1/1
They were then transported in polythene bags to the laboratory, v/v/v/v) were added into the tube. After 10 min at room temperature,
where they were cleaned with filter paper to remove all traces of the tube was dried under vacuum again (about 1 h) and the sample
dust and insects, dried in an air-convection oven at 70°C to was dissolved in a solution containing 95% (v/v) 2 mM Na2HPO4,
constant weight. They were ground in an electric grinder (Blender pH 7.4 and 5% (v/v) acetonitrile.
mill/grater 3), placed in airtight bottles and stored in a dessicator for The analysis was done by reverse phase high performance liquid
analysis. chromatography (HPLC) waters (Millennium), equipped with a
photodiode array detector (Waters model 596), on a column
Picotag C18 (4 mm x 15 cm, Waters). The HPLC column was
Extraction of the oils and collection of the defatted flour equilibrated with buffer A (94% (v/v) 0.14 M CH3COONa containing
3.6 mM triethylamine, pH 6.4, 6% (v/v) acetonitrile) and the elution
Part of the seeds were defatted while the rest were analysed was performed with a convex gradient from 100% buffer A to 46%
undefatted. Oils were extracted from the ground seeds by buffer B (40% H2O/60% acetonitrile, v/v) for 10 min, at a flow rate of
continuous extraction in a Soxhlet apparatus for 8 h using hexane 1 ml/min. Both the column and buffers were maintained at 38°C and
as solvent (AOAC, 1980). The hexane was evaporated on a the absorbance was recorded at 254 nm. An amino acid mixture
rotatory evaporator (Laborata 4003-digital Heidalph) and the (Pierce) of known concentration, derived under the same conditions
defatted cake obtained was dried in an air-convection oven at 60°C was used to identify and to calculate the amino acid concentration.
for 24 h to remove all traces of solvent. This cake was then ground Different concentrations of standard were injected to make a
into flour in a grinder, put in airtight bottles and used for analyses. calibration curve (250, 500, 750 and 1000 picomoles).

Total protein content of defatted seeds: This was calculated from Uncorrected amino acid scores (AAS): This was calculated from
the following formula: the following formula:

Total protein content of defatted flour = B x 100/100-A (g AAS = Essential amino acid in test protein / essential amino acid in
proteins/100 g defatted cake) reference pattern

The reference pattern was the 1985 FAO/WHO 2 to 5 year old

Where, A = lipid content of whole seed and B = protein content of
requirement pattern.
whole seed (Achu et al., 2005).
Protein digestibility corrected amino acid score (PDCAAS):
Soluble protein content of defatted seeds: Most of the soluble This was calculated from the following formula:
nitrogen (non-protein nitrogen) concerns the products of
degradation of proteins such as small peptides and free amino PDCAAS = AAS x true digestibility (TD) (Achu, 2006)
acids (Laure et al., 1971). Soluble nitrogen was extracted from the
defatted samples following the method described by Nilo Rivas et
al. (1981), which was done by precipitating the proteins with a Content in total phenolic compounds of whole seeds
solution of trichloroacetic acid (TCA) and filtration, to obtain a
solution containing non-protein nitrogen. The quantity of nitrogen in The content in total phenolic compounds was determined by the
this solution was determined through digestion of this solution and spectrophotometric method of Marigo (1973). This method uses the
assay by a spectrophotometric method described by Devani et al. Folin-Coicalteu reagent, which is a mixture of phosphotungstic acid
(1989). Ammonium sulphate solution was used as standard and the and phosphomolybdic acid. Through reduction during the oxidation
absorbance was read at 412 nm against the reagent blank. The of phenols, a mixture of tungstene blue and molybdenum oxides is
soluble protein (non-protein nitrogen) content was obtained by formed. Gallic acid solution was used as standard and the
multiplying the soluble nitrogen content by the coefficient 6.25. absorbance of the blue colour was read at 745 nm against the
reagent blank.
True protein content of defatted seeds: This was calculated from
the following formula: Statistical analysis

True protein content = total proteins - soluble proteins (g The data was analyzed using the SPSS 9.0 software. Analysis of
proteins/100 g defatted flour). variance (ANOVA) was used to find the correlation between the
738 Afr. J. Biotechnol.

parameters measured and the different ecological regions of seeds. C. mannii from these two regions has comparable
cultivation of the seeds, and between these parameters in the soluble protein proportions (67 and 64%) as well as C.
different species of seeds. Where the ANOVA test indicated
significant differences, the Student-Newman-Keuls (S-N-K) test was
sativus (94 and 85%). Also for C. sativus, the South West
used to locate these differences. The tests were done at the 5% and the Littoral regions which are located in the Swamp
level of significance. Forest with about the same climate have comparable
proportions (94 and 86%). However, even if the total
protein contents do not vary considerably according to
RESULTS AND DISCUSSION the region (Achu et al., 2005), the profile of these proteins
indicates that they depend on the region: Hot and humid
Total, soluble and true protein contents of defatted regions have higher values of soluble proteins than the
seeds other regions. In this respect, the lower value of 49% for
L. siceraria in the far North might be due to the hot and
The total, soluble and true protein contents of the dry climate of this region. However, the comparable value
defatted seeds are shown in Table 1. The total protein for this sample from the Littoral (46%) with a hot and
contents range from 61.91 (C. sativus) to 73.59% (C. humid climate explains the fact that not only heat and
mannii), soluble proteins from 29.19 (C. maxima) to humidity are responsible for the hydrolysis of proteins in
53.76% (C. sativus) and true proteins from 8.15 (C. the seeds after harvest, but the modes of treatment can
sativus) to 39.53% (C. maxima). There is a significant also influence proteolysis.
difference between the total, soluble and true protein Looking at the various postharvest modes of treatment
contents of the various species of seeds. The total up to the end of drying, it was observed that for some of
protein level of C. mannii is similar to that of C. maxima these seeds, the fruits are usually opened and left for
(68.72%) and L. siceraria (68.52%) but significantly about 2 to 3 days for the pulp to slightly ferment before
higher (p < 0.05) than that of the other seeds, which have the seeds are extracted from the fruit. This is in order to
similar levels. These seeds contain more soluble proteins reduce the sticky and slimy nature of the content of the
than true proteins, except C. maxima where the quantity fruits for easy washing of the seeds. After washing, the
of true proteins is more than that of soluble proteins. C. seeds are dried under the sun. If the sun is not hot
sativus has the highest amount of soluble proteins enough, this drying can take a number of days before the
(53.76%) and the least amount of true proteins (8.15%). seeds are completely dried. This slow drying causes the
This soluble protein level of C. sativus is significantly seeds to remain damp, leading to slow fermentation.
higher (p < 0.05) and the true protein levels of C. mannii Fermentation favours the action of lipolytic enzymes,
and C. maxima are also significantly higher than those of which hydrolyzes triglycerides in the seeds, liberating free
the other seeds. fatty acids (Fokou et al., 2009). This acidity favours the
A view of the various regions of collection of these action of proteases in the seeds which hydrolyze proteins
seeds shows that in all the species, the samples from the into small peptides and free amino acids, leading to an
West have higher proportions of soluble proteins than increase in soluble proteins and a decrease in the true
those from the North West. Samples from the Adamawa protein values. On the other hand, C. maxima whose
region also have high soluble protein values similar to edible fruit (the pumpkin) serves as a delicacy, is not
those from the West. For C. mannii, the total proteins of allowed to ferment before seed extraction. This reduces
this specie from Adamawa have 67% of soluble proteins, the activity of proteases. The seeds are extracted once
against 38% from the South. The West and Adamawa the fruit is mature and the fruit cooked and consumed
regions are in the High Savanna area with similar climate. both by man and animals. This could partly explain the
This area is further away from the sea (inland) than the high true protein values observed in this specie. The fruit
Rain and Swamp forest areas and it receives less rain of C. moschata is also edible, but the seeds once
than the coastal belt. It has good drainage and lacks extracted are usually stored for further planting during the
enough moisture. Rainfall varies depending on the next farming season. Sometimes, they are used for
location of the area to the rain-bearing winds. The climate consumption, but this is only done during periods of
is the equatorial type. In Adamawa, the high elevations in famine, for the seeds are very small in size and so very
this region lend it a relatively cool climate, between 22 to laborious to decorticate, since it is done manually. It is
25°C. Rainfall is within 150 to 200 cm with a long dry also the case with L. siceraria seeds. They are bigger in
period followed by a long wet period. The West region size, but the cork is hard, and so difficult to decorticate.
has moderate to high humidity, the temperature is about Consequently, these seeds are usually stored for long,
22°C, and rainfall is moderated by the mountains and and at room temperature. This may favour the action of
averages 100 to 200 cm per year (Fokou et al., 2009). If proteases, thereby leading to high soluble protein values.
the climate difference could explain this variation in Depending on the water activity (Aw) of the seeds during
values, the high temperature of the Adamawa should be storage, (if it is ≥ 0.6), microbes can set in. Umoh and
more favourable for the activity of proteases, contributing fields (1981) and Collar et al. (1991) showed that during
to this rise in soluble protein values during drying of the fermentation, proteins can be hydrolyzed into volatile
 Achu et al. 739

Table 1. Total, soluble, true protein and phenolic compound contents of seeds.

1 0
*Total Soluble Percentage Phenolic
True proteins
Sample Area Region proteins proteins of soluble compounds
proteins (%)
(%) (%) (%)
C. mannii High Savanna North West 78.38 39.56 50 38.82 0.41
High Savanna West 68.60 43.64 64 24.96 0.30
High Savanna Adamawa 70.0 47.05 67 22.94 0.46
Rain Forest South 69.41 26.18 38 43.23 0.49
Rain Forest East 74.50 31.27 42 43.24 0.32
Swamp Forest South West 80.71 38.19 47 42.52 0.34
Average±SD 73.59± 4.65a 37.65± 7.75c 51 ± 12 35.95 ± 9.46a 0.39  0.08

C. maxima High Savanna North West 71.65 18.08 25 53.57 0.46

High Savanna West 69.08 42.14 61 26.94 0.33
Rain Forest Centre 65.42 27.34 42 38.08 0.46
Average±SD 68.72 ± 2.56ab 29.19 ± 12.14d 43 ± 18 39.53 ± 13.37a 0.42  0.08

C. moschata High Savanna North West 71.46 31.89 45 39.57 0.44

High Savanna West 62.48 33.14 53 29.34 0.52
Rain Forest Centre 61.76 58.33 94 3.43 0.44
Swamp Forest South West 65.66 37.75 57 27.91 0.27
Average±SD 65.34 ± 3.83b 40.28 ± 12.3bc 62 ± 22 25.06± 15.33b 0.42 0.11

L. siceraria Sahel Far North 68.18 33.11 49 35.07 0.29

Savanna North West 65.31 46.77 72 18.54 0.39
High Savanna West 67.98 57.27 84 10.71 0.37
High Savanna Littoral 72.59 33.35 46 39.24 0.32
Swamp Forest Average±SD 68.52± 2.61ab 42.63 ± 11.67b 63 ± 18 25.89 ± 13.50b 0.34  0.05

C. sativus High Savanna West 58.73 49.82 85 8.91 0.54

High Savanna Adamawa 61.20 57.25 94 3.95 0.30
Rain Forest Centre 63.94 49.12 77 14.82 0.53
Swamp Forest South West 59.74 56.03 94 3.71 0.41
Swamp Forest Littoral 65.94 56.56 86 9.38 0.37
b a c
Average±SD 61.91 ±2.67 53.76 ± 3.94 86± 7 8.15 ± 4.58 0.43 0.10
0
= There is no significant difference between the phenolic compound levels of the same specie of seeds from different ecological regions and between
1=
these values in the different species of seeds. *= values are from Achu et al. (2005). Soluble Proteins/Total Proteins x100.SD = Standard deviation.
Each value is a mean of 3 replications.

compounds such as ammonia into the environment as a 73.3% for C. moschata (pumpkin). Lazos (1992) also
result of proteolytic microorganisms such as Clostridia showed that the defatted seed flours of C. pepo and C.
spp. Hence, the various treatments applied to the seeds, maxima have potential food use because of their protein
the duration of storage and drying of the seeds can content of 61.4%. This value is within our range of
influence the soluble protein values. values. The total nitrogen content of the defatted cake of
On the whole, the results show that these “egusi” seeds Coula edulis is 2.1 to 2.4 (Tchiégang et al., 1998), giving
are very rich in proteins especially those of C. mannii. a protein level of 13.12 to 15%, which is much lower than
The total protein levels are similar to those obtained for the values for “egusi” seeds.
defatted Cucurbit seeds by other researchers such as Lal
et al. (1983) who showed values of 60 to 70% for 15
species of Cucurbitaceae seeds from India; Sharma et al. Phenolic compounds
(1986) who found 62.1% for defatted C. melo (musk
melon), 76.1% for Citrullus vulgaris (water melon) and The content in the total phenolic compounds of the
740 Afr. J. Biotechnol.

Table 2. Amino acid composition of defatted C. sativus and C. mannii flours compared to that of defatted soybean meal and casein.

C. sativus C. mannii Soybean meal Casein

1
Amino acid (mg/g of (mg/g of (mg/g of (mg/g of (mg/g ²(mg/g of ²(mg/g of
flour) protein) flour) protein) of flour) protein) protein)
*Histidine 23.6 36.91 27.8 37.32 15.8 26 16.7
Threonine 28.9 45.2 36.6 49.13 18.3 37 47.7
Valine 30 46.92 34.7 46.58 21.7 50 35.5
Isoleucine 29.9 46.76 33.7 45.23 21.2 49 36.8
Leucine 49.2 76.95 55.9 75.03 37.1 82 59.8
**Phenylalanine and Tyrosine 66.9 104.63 70.6 94.77 39.4 90 67.8
Lysine 25.7 40.19 23.4 31.41 32.5 63 -
Total essential amino acids 254.2 397.56 282.7 379.46 186 397 264.3
Aspartic acid 67.3 105.25 80.6 108.19 51.5 116 130
Glutamic acid 133.9 209.42 154.0 206.71 81 191 253.9
Serine 39.2 61.31 44.3 59.46 24.2 52 48.5
Glycine 44.5 69.6 46 61.74 21.4 42 100.8
Arginine 122.4 191.43 128.1 171.95 40 76 20.9
Alanine 36.4 56.93 42.5 57.05 20.8 43 38.9
Proline 25.9 40.51 29.6 39.73 25.2 51 14.7
Total non essential amino acids 469.60 734.45 525.10 704.83 264.10 571.00 607.70
Total amino acids 723.8 1132.01 807.8 1084.3 450.1 968 872

Essential amino acids/ Total

0.35 0.35 0.35 0.35 0.41 0.41 0.3
amino acids (A1)
1 2
Essential amino acids are shown in bold. = Values are from Nwokolo and Sim (1987); = Values are from Pc Priya Chemicals; * = Necessary
only for infants, ** Necessary for synthesis of Tyrosine; - = Not found.

“egusi” seeds ranges from 0.34 (L. siceraria) to 0.43% (C. composition of the different types of phenolic compounds
sativus) (Table 1). The phenolic compound levels of the present in “egusi” seeds is needed in order to confirm
seeds do not depend on the region of cultivation. There is these suggestions.
also no significant difference between the phenolic
compound levels of the different species of seeds. These Amino acid composition of defatted seeds
values are higher than that of pumpkin seed cake (C.
pepo, 0.26%) (Zdunczyk et al., 1999). They are lower The amino acid composition of defatted C. sativus and C.
than those of Canarium schweinfurthii (0.47 to 0.77%) mannii flours are compared to those of defatted soybean
(Kapchie Noutchogoué, 2000) and berries (contain much meal and casein as shown in Table 2. The values are
higher levels of antioxidants than other commonly given in terms of mg/g of flour and protein. The content of
consumed dietary plants), which range from 0.3 to 0.7% non essential amino acids in these flours is more than
of anthocyanins (Törrönen, 2003). Phenolic compounds that of essential amino acids, almost 2 times more. Apart
are natural antioxidants with heart protecting properties. from the combination of phenylalanine and tyrosine,
Those found in nuts and red wine (resveratrol) have leucine is the most abundant essential amino acid while
antioxidant, antithrombotic and anti-inflammatory effects, glutamic acid is the most abundant non essential amino
and inhibit carcinogenesis (Kris-Ertherton et al., 2002). acid in these flours, followed by arginine and aspartic
The total phenolic compounds in these “egusi” seeds can acid (in mg/g of protein). These results are similar to
therefore have beneficial effects on health, though at those of Jacks (1986) who showed that Cucurbit seeds
lower levels than those of walnut, pomegranates and are rich in arginine, aspartic and glutamic acids; and to
berries with high levels of phenolic compounds those of Ojieh et al. (2008) who showed that C. lanatus
(Törrönen, 2003). has good quantities (mg/g of protein) of arginine (90),
These phenolic compounds can thus help to reduce the isoleucine (48), leucine (42), and phenylalanine (32)
atherogenic risk in “egusi” seeds. On the other hand, which are essential amino acids as well as glutamic acid
these phenolic compounds (tannins) can bind proteins, (169) and aspartic acid (163) and that “egusi” melon
carbohydrates, fats, and minerals, making them compares favourably with the known protein rich foods
unavailable (Ganora, 2005). However, a study of the such as soybean, cowpeas, pigeon peas and pumpkin.
 Achu et al. 741

Table 3. Essential amino acids and uncorrected and corrected amino acid scores.

3
C. sativus C. mannii Reference
Amino Acid (mg/g of 1 2 (mg/g of protein
AAS PDCAAS AAS PDCAAS
protein) protein) (mg/g of protein)
Histidine 36.91 1.94 1.87 37.32 1.96 1.74 19
Threonine 45.2 1.33 1.28 49.13 1.44 1.28 34
Valine 46.92 1.34 1.29 46.58 1.33 1.18 35
Isoleucine 46.76 1.67 1.61 45.23 1.62 1.43 28
Leucine 76.95 1.17 1.12 75.03 1.14 1.01 66
Phenylalanine and Tyrosine 104.63 1.03 0.99 94.77 0.87 0.77 63
Lysine 40.19 0.69 0.67 31.41 0.54 0.48 58
Total essential amino acids 397.56 379.46 303
1 2
AAS: Amino acid score (uncorrected) = essential amino acid in test protein / essential amino acid in reference protein; PDCAAS: protein
3 4
digestibility corrected amino acid score = AAS x true digestibility (TD); 1985 FAO/WHO 2 to 5-year old requirement pattern; True digestibility (TD):
TD = 96.19% (C. sativus) and 88.7% (C. mannii) (Achu, 2006).

C. sativus has higher levels of essential amino acids than at low levels of inclusion), the high carbohydrate-low
C. mannii. These “egusi” seeds also have higher levels of protein diets of people in the tropics). However, a blend
total essential amino acids than casein. The amino acid of “egusi” and soybean meal will greatly improve the
composition of the egusi seeds therefore shows that they nutritional value of these seeds when used in preparing
have an excellent protein configuration. They have higher sauces.
levels of histidine, threonine, phenylalanine and tyrosine Jacks (1986) and Martin (1998) showed that the
than soybean, which has higher levels of lysine, leucine, proteins of Cucurbit seeds are mostly of globulin type.
isoleucine and valine. The total level of amino acids in They are deficient in lysine and in sulphur-containing
terms of mg/g of protein (essential and non essential amino acids. These are similar to the results obtained in
amino acids) is higher in C. sativus than in C. mannii, these seeds. The results of these “egusi” seeds are also
soybean and casein, while the A1 ratio (A1 = sum of similar to those of C. Pepo seed cake with respect to
essential amino acids/total amino acids) is higher in lysine. The proteins of pumpkin seed cake (C. pepo)
soybean (0.41) than in C. sativus ( 0.35), C. mannii (0.35) contain low levels of lysine (3.21 g/16 g N or 32.1 mg/g of
and casein (0.3). The essential amino acid values of protein) and isoleucine (38.3 mg/g of protein) (Zdunczyk
these seeds are generally higher than those of melon and et al., 1999). These “egusi” seeds have higher levels of
fluted pumpkin seed meals. These seed flours are isoleucine than those of pumpkin seed cake (Zdunczyk et
superior to soybean in their content of all amino acids al., 1999). The results are similar to those of C. sativus
except lysine, when the amino acids are measured in and L. vulgaris seeds, in which lysine was shown to be
terms of mg/g of flour (Nwokolo and Sim, 1987). This the first limiting amino acid (Kanar et al., 2006).
means that in the absence of tryptophan, lysine is the All these results together with those of these “egusi”
limiting amino acid in these seeds. However, looking at seeds showed that lysine is the limiting amino acid in
the amino acid values in terms of mg/g of protein, these these seeds. The results are different from those of
“egusi” flours are inferior to soybean in their essential Mansour et al. (1993a) who showed that isoleucine and
amino acid levels except histidine (essential for infants) valine were the first limiting amino acids for C. pepo seed
threonine, phenylalanine and tyrosine. These seeds also meal, and those of the African pear cake (Dacryodes
have higher levels of valine, isoleucine, leucine, edulis) by Omoti and Okiy (1987) who showed that this
phenylalanine and tyrosine than casein. This shows that fruit contains high contents of the essential amino acids,
they can be good for infants. They have higher levels of lysine, leucine and threonine. The “egusi” seeds studied
most non essential amino acids than soybean except also contain high contents of leucine.
aspartic acid and proline. These give “egusi” meals a
lower ratio of A1 (0.35) than soybean (0.41). The higher
A1 ratio in soybean than these seeds means that soybean Essential amino acids, uncorrected and corrected
generally has higher levels of essential amino acids than amino acid scores of “egusi” seed flours
these “egusi” seed meals. However, the levels of
essential and other amino acids of these seeds are high The uncorrected AAS and PDCAAS of C. sativus and C.
enough (for some of their essential amino acid levels are mannii seed flours are shown in Table 3. The results
higher than that of the 2 to 5 years old reference protein show that the PDCAAS of C. sativus is 0.67 and that of
(Table 3) to enable them to adequately supplement (even C. mannii is 0.48. These values are those of lysine,
742 Afr. J. Biotechnol.

indicating that in the absence of methionine and Cameroon. Thesis presented and defended for the award of a
Doctorate PhD in Biochemistry. Faculty of Science, University of
tryptophan, lysine is the limiting amino acid in these
Yaounde I. p. 180
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have higher levels of the essential amino acids, histidine
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tyrosine than soybean and higher levels of valine, Technol. 2(1):63-68.
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IA (2007). Macronutrient composition of three cucurbit species
Part of this research was supported by the International cultivated for seed consumption in Côte d’Ivoire. Afr. J. Biotechnol.
6(5):529-533.
Foundation for Science, Stockholm, Sweden and The
Mariod AA, Ahmed YM, Matthäus B, Khaleel G, Siddig A, Gabra AM,
Organization for the Prohibition of Chemical Weapons Abdelwahab SI (2009). A comparative study of the properties of six
(OPCW), THE HAGUE, Netherlands, through a grant to Sudanese cucurbit seeds and seed oils. J. Am. Oil Chem. Soc.
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